The invention relates to novel aldehyde dehydrogenase-like sequences and proteins. Also provided are vectors, host cells, and recombinant methods for making and using the novel molecules.
Aldehydes are a class of highly-reactive molecules that are involved in a number of biological processes. The biological effects mediated by some aldehydes are beneficial, but other members of this class of molecule are associated with biological toxicity, mutagenesis, and carcinogenesis. Aldehydes are generated from a large number of endogenous and exogenous sources. The metabolism of amino acids, lipids, steroids, vitamins, and some amines leads to the formation of aldehydes, and they can also be derived from a number of drugs and environmental agents (reviewed by Lindahl, (1992) Crit. Rev. Biochem. Mol. Biol. 27:283-335).
The aldehyde dehydrogenases make up a large family of enzymes that oxidize aldehydes to their less-reactive carboxylic acids. The members of this family of NAD+ (nicotinamide adenine dinucleotide)- or NADP (nicotinamide adenine dinucleotide phosphate)-dependent enzymes share structural and functional features and generally exhibit broad substrate specificity, catalyzing the oxidation of both aliphatic and aromatic aldehydes (reviewed by Lindahl, supra). The aldehyde dehydrogenase family includes both cytoplasmic and mitochondrial members. Various members of the mammalian family of aldehyde dehydrogenase are expressed in liver, stomach, lung, kidney, testis, salivary gland, muscle, heart, and brain (reviewed by Yoshida et al., (1998) Eur. J. Biochem. 251:549-557).
Aldehyde dehydrogenases are also involved in a number of key metabolic pathways, including, but not limited to, the metabolism of arginine, proline, glutamate, glycine, serine, threonine, histidine, tyrosine, tryptophan, ascorbate, aldarate, xcex2-alanine, butanoate, fatty acids, glycerolipids, pyruvate, propanoate, 4-aminobutyric acid (GABA), retinoic acid, and xenobiotics; the degradation of lysine, valine, leucine, and isoleucine; and the biosynthesis of bile acids. In addition, mutations in members of the human aldehyde dehydrogenase have been associated with a number of diseases including Sjxc3x6gren-Larsson syndrome, alcohol flushing syndrome, and type II hyperprolinemia (reviewed by Vasiliou et al., (1999) Pharmacogenetics 9: 421-434). Changes in aldehyde dehydrogenase activity have also been associated with a number of cancers, including liver, urinary bladder, colon, and mammary cancers (reviewed by Lindahl, (1992) Crit. Rev. Biochem. Mol. Biol. 27:283-335).
The critical role that aldehyde dehydrogenases play in numerous biological processes and diseases make them important targets for therapeutic intervention. It is consequently important to identify novel genes coding for members of this enzyme family.
The present invention is based, in part, on the discovery of a novel human aldehyde dehydrogenase, referred to herein as xe2x80x9c32140xe2x80x9d. The nucleotide sequence of a cDNA encoding 32140 is shown in SEQ ID NO:1, and the amino acid sequence of a 32140 polypeptide is shown in SEQ ID NO:2. In addition, the nucleotide sequence of the coding region is depicted in SEQ ID NO:3.
Accordingly, in one aspect, the invention features a nucleic acid molecule which encodes a 32140 protein or polypeptide, e.g., a biologically active portion of the 32140 protein. In a preferred embodiment, the isolated nucleic acid molecule encodes a polypeptide having the amino acid sequence of SEQ ID NO:2. In other embodiments, the invention provides an isolated 32140 nucleic acid molecule having the nucleotide sequence shown in SEQ ID NO:1, SEQ ID NO:3, or the sequence of the DNA insert of the plasmid deposited with ATCC as Accession Number PTA-3424. In still other embodiments, the invention provides nucleic acid molecules that are substantially identical (e.g., naturally occurring allelic variants) to the nucleotide sequence shown in SEQ ID NO: 1, SEQ ID NO:3, or the sequence of the DNA insert of the plasmid deposited with ATCC as Accession Number PTA-3424. In other embodiments, the invention provides a nucleic acid molecule which hybridizes under stringent hybridization conditions to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, or the sequence of the DNA insert of the plasmid deposited with ATCC as Accession Number PTA-3424, wherein the nucleic acid encodes a full length 32140 protein or an active fragment thereof.
In a related aspect, the invention further provides nucleic acid constructs, which include a 32140 nucleic acid molecule described herein. In certain embodiments, the nucleic acid molecules of the invention are operatively linked to native or heterologous regulatory sequences. Also included are vectors and host cells containing the 32140 nucleic acid molecules of the invention e.g., vectors and host cells suitable for producing 32140 nucleic acid molecules and polypeptides.
In another related aspect, the invention provides nucleic acid fragments suitable as primers or hybridization probes for the detection of 32140-encoding nucleic acids.
In still another related aspect, isolated nucleic acid molecules that are antisense to a 32140 encoding nucleic acid molecule are provided.
In another aspect, the invention features 32140 polypeptides, and biologically active or antigenic fragments thereof that are useful, e.g., as reagents or targets in assays applicable to treatment and diagnosis of 32140-mediated or -related disorders. In another embodiment, the invention provides 32140 polypeptides having a 32140 activity. Preferred polypeptides are 32140 proteins including at least one aldehyde dehydrogenase domain, and, preferably, having a 32140 activity, e.g., a 32140 activity as described herein.
In other embodiments, the invention provides 32140 polypeptides, e.g., a 32140 polypeptide having the amino acid sequence shown in SEQ ID NO:2; the amino acid sequence encoded by the cDNA insert of the plasmid deposited with ATCC as Accession Number PTA-3424; an amino acid sequence that is substantially identical to the amino acid sequence shown in SEQ ID NO:2; or an amino acid sequence encoded by a nucleic acid molecule having a nucleotide sequence which hybridizes under stringent hybridization conditions to a nucleic acid molecule comprising the nucleotide sequence of SEQ ID NO:1, SEQ ID NO:3, or the sequence of the DNA insert of the plasmid deposited with ATCC as Accession Number PTA-3424, wherein the nucleic acid encodes a full length 32140 protein or an active fragment thereof.
In a related aspect, the invention further provides nucleic acid constructs which include a 32140 nucleic acid molecule described herein.
In a related aspect, the invention provides 32140 polypeptides or fragments operatively linked to non-32140 polypeptides to form fusion proteins.
In another aspect, the invention features antibodies and antigen-binding fragments thereof, that react with, or more preferably specifically bind 32140 polypeptides.
In another aspect, the invention provides methods of screening for compounds that modulate the expression or activity of the 32140 polypeptides or nucleic acids.
In still another aspect, the invention provides a process for modulating 32140 polypeptide or nucleic acid expression or activity, e.g. using the screened compounds. In certain embodiments, the methods involve treatment of conditions related to aberrant activity or expression of the 32140 polypeptides or nucleic acids, such as conditions involving aberrant or deficient cellular proliferation or differentiation.
More particularly, 32140 is associated with viral disease. It is induced in Herpes Simplex Virus (HSV)-infected human ganglia and neuroblastoma. It was observed to be induced up to 5-fold in HSV-infected neuroblastoma cells. It is hypothesized that HSV induces expression of a novel 10-formyltetrahydrofolate DH isozyme encoded by gene 32140, particularly in infected neurons.
The invention also provides assays for determining the activity of or the presence or absence of 32140 polypeptides or nucleic acid molecules in a biological sample, including assays for the diagnosis of disease. Gene 32140 showed expression in the following tissues, including but not limited to: brain, spinal cord, heart, mammary gland, salivary gland, small intestine, stomach, testis, uterus, and trachea.
In further aspect the invention .provides assays for determining the presence or absence of a genetic alteration in a 32140 polypeptide or nucleic acid molecule, including assays for the diagnosis of disease.